Clonal propagation of primate offspring by embryo splitting

• US 20030106082A1
• Filed: 01/09/2003
• Published: 06/05/2003
• Est. Priority Date: 01/07/2000
• Status: Abandoned Application

• Current Assignee: Emory University, Magee-Womens Health Corporation
• Original Assignee: Oregon Health & Science University
• Inventors: Chan, Anthony W.S., Schatten, Gerald
• Application Number:
  US10/340,332
• Publication Number:
  US 20030106082A1
• Time in Patent Office: Days
• US Class Current: 800/14
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/00 Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 15/873 Techniques for producing new embryos, e.g. nuclear transfer, manipulation of totipotent cells or production of chimeric embryos
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696

Abstract

The present invention relates to the clonal propagation of primate offspring by embryo splitting. Here, genetically identical nonhuman embryos may be produced as twin and larger sets by separation and reaggregation of blastomeres of cleavage-stage embryos. Furthermore, the present invention also relates to methods for producing embryonic stem cells and transgenic embryonic stem cells isolated from dissociated blastomeres.

Claims

1. A method for cloning an animal comprising the steps of:

dissociating blastomeres from embryos;

transferring said blastomeres to empty zonae;

culturing said blastomeres to an embryonic stage;

transferring said embryos to the oviducts of surrogate females; and

producing a cloned animal by parturition.

2. The method of claim 1, wherein said animal is selected from the group consisting of mammals, birds, reptiles, amphibians, and fish.

3. The method of claim 2, wherein said animal is a primate.

4. The method of claim 3, wherein said animal is a nonhuman primate.

5. The method of claim 4, wherein said nonhuman primate is a monkey.

6. The method of claim 1, wherein said embryo is at the 4- to 8-cell stage.

7. The method of claim 1, wherein said embryo is transgenic.

8. The method of claim 1, wherein said embryos are frozen and stored prior to said transferring to surrogate females.

9. The method of claim 1, further comprising the step of producing embryonic stem cells from said dissociated blastomeres.

10. The method of claim 7, further comprising the step of producing embryonic stem cells from said dissociated blastomeres.

11. An animal produced according to the method of claim 1.

12. The animal of claim 11, wherein said animal is a primate.

13. The animal of claim 12, wherein said animal is a nonhuman primate.

14. An animal produced according to the method of claim 7.

15. The animal of claim 14, wherein said animal is a primate.

16. The animal of claim 15, wherein said animal is a nonhuman primate.

53. The method of claim 1, further comprising the step of performing preimplantation genetic diagnosis on said embryo.

54. The method of claim 53, wherein said preimplantation genetic diagnosis is performed prior to transfer to the oviduct of a female surrogate.

55. The method of claim 54, wherein said preimplantation genetic diagnosis is selected from the group comprising PCR, FISH, SSCP, RFLP, PRINS, CGH, COMET analysis, heteroduplex analysis, Southern analysis, and DGGE analysis.

59. The method of claim 1, wherein said blastomeres are frozen.

60. The method of claim 59, wherein said blastomeres are stored in a repository.

17. A method of producing embryonic stem cells comprising the steps of:

dissociating blastomeres from embryos; and

culturing said blastomeres to produce stem cell lines.

18. The method of claim 17, wherein said embryonic stem cells are primate embryonic stem cells.

19. The method of claim 18, wherein said primate embryonic stem cells are nonhuman primate embryonic stem cells.

20. The method of claim 17, wherein said embryonic stem cells are transgenic embryonic stem cells.

21. The method of claim 20, wherein said transgenic embryonic stem cells are transgenic primate embryonic stem cells.

22. The method of claim 21, wherein said transgenic primate embryonic stem cells are transgenic nonhuman primate embryonic stem cells.

23. An embryonic stem cell produced according to the method of claim 17.

24. The embryonic stem cell of claim 23, wherein said embryonic stem cell is stored in an embryonic cell repository.

25. The embryonic stem cell of claim 23, wherein said embryonic stem cell is used for gene therapy.

26. The embryonic stem cell of claim 23, wherein said embryonic stem cell is used as a therapy for human disease.

27. The embryonic stem cell of claim 26, wherein said human disease is selected from the group consisting of cardiovascular diseases, neurological diseases, reproductive disorders, cancers, eye diseases, endocrine disorders, pulmonary diseases, metabolic disorders, hereditary diseases, autoimmune disorders, and aging.

28. An embryonic stem cell produced according to the method of claim 18.

29. The embryonic stem cell of claim 28, wherein said embryonic stem cell is stored in an embryonic cell repository.

30. The embryonic stem cell of claim 28, wherein said embryonic stem cell is used for gene therapy.

31. The embryonic stem cell of claim 28, wherein said embryonic stem cell is used as a therapy for human disease.

32. The embryonic stem cell of claim 31, wherein said human disease is selected from the group consisting of cardiovascular diseases, neurological diseases, reproductive disorders, cancers, eye diseases, endocrine disorders, pulmonary diseases, metabolic disorders, hereditary diseases, autoimmune disorders, and aging.

33. An embryonic stem cell produced according to the method of claim 19.

34. The embryonic stem cell of claim 33, wherein said embryonic stem cell is stored in an embryonic cell repository.

35. The embryonic stem cell of claim 33, wherein said embryonic stem cell is used for gene therapy.

36. The embryonic stem cell of claim 33, wherein said embryonic stem cell is used as a therapy for human disease.

37. The embryonic stem cell of claim 36, wherein said human disease is selected from the group consisting of cardiovascular diseases, neurological diseases, reproductive disorders, cancers, eye diseases, endocrine disorders, pulmonary diseases, metabolic disorders, hereditary diseases, autoimmune disorders, and aging.

38. An embryonic stem cell produced according to the method of claim 20.

39. The embryonic stem cell of claim 38, wherein said embryonic stem cell is stored in an embryonic cell repository.

40. The embryonic stem cell of claim 38, wherein said embryonic stem cell is used for gene therapy.

41. The embryonic stem cell of claim 38, wherein said embryonic stem cell is used as a therapy for human disease.

42. The embryonic stem cell of claim 41, wherein said human disease is selected from the group consisting of cardiovascular diseases, neurological diseases, reproductive disorders, cancers, eye diseases, endocrine disorders, pulmonary diseases, metabolic disorders, hereditary diseases, autoimmune disorders, and aging.

43. An embryonic stem cell produced according to the method of claim 21.

44. The embryonic stem cell of claim 43, wherein said embryonic stem cell is stored in an embryonic cell repository.

45. The embryonic stem cell of claim 43, wherein said embryonic stem cell is used for gene therapy.

46. The embryonic stem cell of claim 43, wherein said embryonic stem cell is used as a therapy for human disease.

47. The embryonic stem cell of claim 46, wherein said human disease is selected from the group consisting of cardiovascular diseases, neurological diseases, reproductive disorders, cancers, eye diseases, endocrine disorders, pulmonary diseases, metabolic disorders, hereditary diseases, autoimmune disorders, and aging.

48. An embryonic stem cell produced according to the method of claim 22.

49. The embryonic stem cell of claim 48, wherein said embryonic stem cell is stored in a repository.

50. The embryonic stem cell of claim 48, wherein said embryonic stem cell is used for gene therapy.

51. The embryonic stem cell of claim 48, wherein said embryonic stem cell is used as a therapy for human disease.

52. The embryonic stem cell of claim 51, wherein said human disease is selected from the group consisting of cardiovascular diseases, neurological diseases, reproductive disorders, cancers, eye diseases, endocrine disorders, pulmonary diseases, metabolic disorders, hereditary diseases, autoimmune disorders, and aging.

56. A method for preimplantation genetic diagnosis of an embryo comprising the steps of:

dissociating a blastomere from an embryo; and

performing genetic analysis on said blastomere prior to implantation of said embryo.

57. The method of claim 56, wherein said embryo is implanted into a female surrogate.

58. The method of claim 56, wherein said genetic analysis is selected from the group comprising PCR, FISH, SSCP, RFLP, PRINS, CGH, COMET analysis, heteroduplex analysis, Southern analysis, and DGGE analysis.